Figure 1

LUCL is a multi-copy, single-insertion transgene.(A) LUCL as a multi-copy transgene. Only two tandemcopies are shown, with each copy extending from RB (right border of thetransfer DNA) to LB (left border of the transfer DNA). Restriction sitesand distances between sites are noted. The question mark indicates theunknown distance between two tandemly arrayed copies. The stars indicatethe miR172 binding sites. The red lines depict the region used as aprobe in the Southern blots in (C) and (D). (B)Luciferase luminescence from LUCL and LUCH seedlings.Ten-day-old seedlings grown on the same plate were imaged for luciferaseluminescence using a CCD camera. The blue spots in the LUCHsector represent seedlings with luciferase luminescence. The lack ofsignals in the LUCL sectors represents the absence ofluciferase luminescence. (C) Southern blot analysis ofLUCL, Col-0 and LUCH. The gray triangle indicatesincreasing amounts of genomic DNA from LUCH; the left lane hasan amount of DNA equal to LUCL whereas the right lane containstwice that of LUCL. Genomic DNA was digested withEcoRI and hybridized with a probe corresponding to theLUC coding region (red line in (A)). The 2.1-kb bandcorresponding to the LUC-AP2 fragment is indicated by a redarrow. The intensity of the 2.1-kb band in LUCL is much higherthan that in LUCH. (D) Southern blot analysis ofLUCL and Col-0. Genomic DNA was digested withBamHI and hybridized to a probe corresponding to theLUC coding region (red line in (A)). Theapproximately 6-kb band (red arrow) represents the possibility of amulti-copy transgene as the distance between the two BamHIsites in two tandemly arrayed copies is 5.4 kb (not counting theunknown distance between the LB and RB (question mark)).